Pharmaceutical compositions containing a glycopeptide antibiotic and a cyclodextrin

ABSTRACT

Disclosed are pharmaceutical compositions containing a cyclodextrin and a therapeutically effective amount of a glycopeptide antibiotic or a salt thereof. Also disclosed are methods of treating a bacterial disease in a mammal by administering such pharmaceutical compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.09/846,893, filed May 1, 2001, now U.S. Pat. No. 6,858,584; whichapplication claims the benefit of U.S. Provisional Application No.60/201,178, filed 02 May 2000; and U.S. Provisional Applications Nos.60/213,415; 60/213,410; 60/213,417; 60/213,146; 60/213,428, all filed 22Jun. 2000; and U.S. Provisional Application No. 60/226,727, filed 18Aug. 2000, which applications are incorporated herein by reference intheir entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention is directed to novel pharmaceutical compositionscomprising a cyclodextrin and a therapeutically effective amount of aglycopeptide antibiotic. This invention is also directed to methods fortreating a bacterial disease in a mammal using such pharmaceuticalcompositions.

2. State of the Art

Glycopeptide antibiotics and lipidated derivatives thereof arewell-known in the art (see Glycopeptide Antibiotics, edited by R.Nagarajan, Marcel Dekker, Inc. New York (1994)). These glycopeptidecompounds are highly effective antibiotics for treating a wide varietyof bacterial diseases in mammals. However, when administered to amammal, some glycopeptide antibiotics exhibit undesirable properties,such as excessive tissue accumulation, nephrotoxicity, histamine release(Red Man Syndrome) and vascular irritation. Accordingly, a need existsfor novel pharmaceutical compositions of glycopeptide antibiotics whichreduce these undesired properties.

SUMMARY OF THE INVENTION

The present invention provides novel pharmaceutical compositionscomprising a cyclodextrin and a therapeutically effective amount of aglycopeptide antibiotic. Surprisingly, when administered to a mammal,the pharmaceutical compositions of this invention exhibit one or more ofthe following properties (a) reduced tissue accumulation of theglycopeptide antibiotic, (b) reduced nephrotoxicity, (c) reducedhistamine release (Red Man Syndrome) and (d) reduced vascularirritation, compared to a pharmaceutical composition which does notcontain the cyclodextrin.

By reducing the undesirable effects of the glycopeptide (e.g.nephrotoxicity), administration of the glycopeptide in combination witha cyclodextrin increases the therapeutic window for the glycopeptide,and allows a greater amount to be administered.

Accordingly, in one of its composition aspects, this invention isdirected to a pharmaceutical composition comprising a cyclodextrin and atherapeutically effective amount of a glycopeptide antibiotic, or apharmaceutically acceptable salt thereof. This preferred composition canbe in the form of a lyophilized powder or a sterilized powder.

In another one of its composition aspects, this invention is directed toa pharmaceutical composition comprising an aqueous cyclodextrin solutionand a therapeutically effective amount of a glycopeptide antibiotic, ora pharmaceutically acceptable salt thereof.

In another preferred embodiment, the pharmaceutical compositioncomprises:

-   -   (a) a therapeutically effective amount of a glycopeptide        antibiotic, or a pharmaceutically acceptable salt thereof;    -   (b) 1 to 40 weight percent of a cyclodextrin; and    -   (c) 60 to 99 weight percent of water, provided that the        components of the composition total 100 weight percent.

Preferably, the cyclodextrin employed in the pharmaceutical compositionsof this invention is hydroxypropyl-β-cyclodextrin or sulfobutyl etherβ-cyclodextrin. More preferably, the cyclodextrin ishydroxypropyl-β-cyclodextrin. Preferably, the cyclodextrin will compriseup to about 90 weight percent, and typically about 1 to 40 weightpercent; preferably, about 5 to 35 weight percent; more preferable,about 10 to 30 weight percent, of the formulation.

Preferably, the glycopeptide antibiotic employed in the pharmaceuticalcompositions of this invention is a lipidated glycopeptide antibiotic.Preferably, the glycopeptide antibiotic will be present in thepharmaceutical composition in a therapeutically effective amount.

In a preferred embodiment, the glycopeptide antibiotic employed in thisinvention is a compound of formula I

wherein:

R¹ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic and —R^(a)—Y—R^(b)-(Z)_(x);or R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)-(Z)_(x);

R² is hydrogen or a saccharide group optionally substituted with—R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)-(Z)_(x);

R³ is —OR^(c), —NR^(c)R^(c), —O—R^(a)—Y—R^(b)-(Z)_(x),—NR^(c)—R^(a)—Y—R^(b)-(Z)_(x), —NR^(c)R^(e), or —O—R^(e); or R³ is anitrogen-linked, oxygen-linked, or sulfur-linked substituent thatcomprises one or more phosphono groups;

R⁴ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)-(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)-(Z)_(x);

R⁵ is selected from the group consisting of hydrogen, halo,—CH(R^(c))—NR^(c)R^(c), —CH(R^(c))—NR^(c)R^(e),—CH(R^(c))—NR^(c)—R²—Y—R^(b)-(Z)_(x), —CH(R^(c))—R^(x),—CH(R^(c))—NR^(c)—R^(a)—C(═O)—R^(x), and a substituent that comprisesone or more phosphono groups;

R⁶ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)-(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —NR^(c)—R^(a)—Y—R^(b)-(Z)_(x), or R⁵ and R⁶ can bejoined, together with the atoms to which they are attached, form aheterocyclic ring optionally substituted with—NR^(c)—R^(a)—Y—R^(b)-(Z)_(x);

R⁷ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)-(Z)_(x), and —C(O)R^(d);

R⁸ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R⁹ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R¹⁰ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic; or R⁸ and R¹⁰ arejoined to form —Ar¹—O—Ar²—, where Ar¹ and Ar² are independently aryleneor heteroarylene;

R¹¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic, or R¹⁰ and R¹¹ arejoined, together with the carbon and nitrogen atoms to which they areattached, to form a heterocyclic ring;

R¹² is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic, —C(O)R^(d), —C(NH)R^(d),—C(O)NR^(c)R^(c), —C(O)OR^(d), —C(NH)NR^(c)R^(c) and—R^(a)—Y—R^(b)-(Z)_(x), or R¹¹ and R¹² are joined, together with thenitrogen atom to which they are attached, to form a heterocyclic ring;

R¹³ is selected from the group consisting of hydrogen or —OR¹⁴;

R¹⁴ is selected from hydrogen, —C(O)R^(d) and a saccharide group;

each R^(a) is independently selected from the group consisting ofalkylene, substituted alkylene, alkenylene, substituted alkenylene,alkynylene and substituted alkynylene;

each R^(b) is independently selected from the group consisting of acovalent bond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen;

each R^(c) is independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclicand —C(O)R^(d);

each R^(d) is independently selected from the group consisting of alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

R^(e) is a saccharide group;

each R^(f) is independently alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, or heterocyclic;

R^(x) is an N-linked amino saccharide or an N-linked heterocycle;

X¹, X² and X³ are independently selected from hydrogen or chloro;

each Y is independently selected from the group consisting of oxygen,sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)C(O)—, —OSO₂—, —OC(O)—,—NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂O—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)C(O)NR^(c)—,—OC(O)NR^(c)—, —C(═O)—, and —NR^(c)SO₂NR^(c)—;

each Z is independently selected from hydrogen, aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic;

n is 0, 1 or 2; and

x is 1 or 2;

or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

Preferably, R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or —C(O)—R^(a)—Y—R^(b)-(Z).More preferably R¹ is an amino saccharide group substituted on thesaccharide nitrogen with —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃;—CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph; —CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—NH—CH₂-4-[(4—(CH₃)₂CHCH₂—]-Ph; —CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph;—CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph; or —CH₂-4-(4-Cl-Ph)-Ph. Morepreferably R¹ can also be an amino saccharide group substituted on thesaccharide nitrogen with 4-(4-chlorophenyl)benzyl or4-(4-chlorobenzyloxy)benzyl.

More preferably, R¹ is a saccharide group of the formula:

wherein R¹⁵ is —R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)-(Z)_(x); and R¹⁶ is hydrogen or methyl.

Preferably, R¹⁵ is —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃;—CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph; —CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph; —CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph;—CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph; or —CH₂-4-(4-Cl-Ph)-Ph.

Preferably, R¹⁵ can also be 4-(4-chlorophenyl)benzyl or4-(4-chlorobenzyloxy)benzyl.

Preferably, R² is hydrogen.

Preferably, R³ is —OR^(c); —NR^(c)R^(c). More preferably, R³ is —OH;—NH—(CH₂)₃—N(CH₃)₂; N-(D-glucosamine); —NHCH(CO₂CH₃)CH₂CO₂CH₃;—NH(CH₂)₃-(morpholin-4-yl); —NH(CH₂)₃—NH(CH₂)₂CH₃ ;—NH(CH₂-piperidin-1-yl; —NH(CH₂)₄NHC(N)NH₂; —NH(CH₂)₂—N⁺(CH₃)₃;—NHCH(COOH)(CH₂)₃NHC(N)NH₂; —NH—[CH₂CH₂CH₂—NH—]₃—H; —N[(CH₂)₃N(CH₃)₂]₂;—NH(CH₂)₃-imidazol-1-yl; —NHCH₂-4-pyridyl; —NH(CH₂)₃CH₃; —NH(CH₂)₂OH;—NH(CH₂)₅OH; —NH(CH₂)₂OCH₃; —NHCH₂-tetrahydrofuran-2-yl; —N[(CH₂)₂OH]₂;—NH(CH₂)₂N[(CH₂)₂OH]₂; —NHCH₂COOH; —NHCH(COOH)CH₂OH; —NH(CH₂)₂COOH;N-(glucamine); —NH(CH₂)₂COOH; —NH(CH₂)₃SO₃H; —NHCH(COOH)(CH₂)₂NH₂;—NHCH(COOH)(CH₂)₃NH₂; —NHCH(COOH)CH₂CO₂(CH₂)₃—N⁺(CH₃)₃;—NHCH(COOH)CH₂CO₂(CH₂)₂C(O)—N(CH₃)₂;—NHCH(COOH)CH₂CO₂(CH₂)₃-morpholin-4-yl;—NHCH(COOH)CH₂CO₂(CH₂)₂OC(O)C(CH₃)₃; —NHCH(CH₂COOH)CO₂(CH₂)₃—N⁺(CH₃)₃;—NHCH(CH₂COOH)CO₂(CH₂)₂C(O)N(CH₃)₂;—NHCH(CH₂COOH)CO₂(CH₂)₃-morpholin-4-yl;—NHCH(CH₂COOH)CO₂(CH₂)₂OC(O)C(CH₃)₃; —NHCH(COOH)CH₂CO₂CH₃;—NHCH(CH₂COOH)CO₂(CH₂)₂N(CH₃)₂; —NHCH(COOH)CH₂CO₂CH₂C(O)N(CH₃)₂;—NHCH(CH₂COOH)CO₂CH₂C(O)N(CH₃)₂; —NHCH(CH₂COOH)CO₂CH₃; —NH(CH₂)₃N(CH₃)₂;—NHCH₂CH₂CO₂CH₃; —NHCH[CH₂CO₂CH₂C(O)N(CH₃)₂]CO₂CH₂—C(O)—N(CH₃)₂;—NHCH₂CO₂CH₃; —N-(methyl3-amino-3-deoxyaminopyranoside);—N-(methyl3-amino-2,3,6-trideoxyhexopyranoside);—N-(2-amino-2-deoxy-6-(dihydrogenphosphate)glucopyranose;—N-(2-amino-2-deoxygluconic acid); —NH(CH₂)₄COOH; —N—(N—CH₃-D-glucamine;—NH(CH₂)₆COOH; —O(D-glucose); —NH(CH₂)₃OC(O)CH(NH₂)CH₃;—NH(CH₂)₄CH(C(O)-2-HOOC-pyrrolidin-1-yl)NHCH(COOH)—CH₂CH₂Ph (S,Sisomer); —NH—CH₂CH₂—NH—(CH₂)₉CH₃; —NH(CH₂)C(O)CH₂C(O)N(CH₃)₂. Still morepreferably, R³ is —OH.

Preferably, R⁴, R⁶ and R⁷ are each independently selected from hydrogenor —C(O)R^(d). More preferably, R⁴, R⁶ and R⁷ are each hydrogen.

Preferably, R⁵ is hydrogen, —CH₂—NHR^(c), —CH₂—NR^(c)R^(e),—CH₂—NH—R^(a)—Y—R^(b)-(Z)_(x), or a substituent comprising one or twophosphono groups. When R⁵ is phosphono-containing substituent, R⁵ ispreferably a group of the formula —CH₂—NH—R^(a)—P(O)(OH)₂, where R^(a)is as defined herein. In this formula, R^(a) is preferably an alkylenegroup. Particularly preferred R⁵ substituents includeN-(phoshonomethyl)aminomethyl;N-(2-hydroxy-2-phosphonoethyl)aminomethyl;N-carboxymethyl-N-(2-phosphonoethyl)aminomethyl;N,N-bis(phosphonomethyl)aminomethyl; N-(3-phosphonopropyl)aminomethyl;and the like.

Preferably, when R⁵ is not a phosphono-containing substituent, R⁵ ishydrogen, —CH₂—NHR^(c), —CH₂—NR^(c)R^(e) or—CH₂—NH—R^(a)—Y—R^(b)-(Z)_(x). R⁵ can also preferably be hydrogen;—CH₂—N—(N—CH₃-D-glucamine); —CH₂—NH—CH₂CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—NHC(O)—(CH₂)₃COOH; —CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—COOH; —CH₂—NH—(CH₂)₅COOH; —CH₂-(morpholin-4-yl);—CH₂—NH—CH₂CH₂—O—CH₂CH₂OH; —CH₂—NH—CH₂CH(OH)—CH₂OH; —CH₂—N[CH₂CH₂OH]₂;—CH₂—NH—(CH₂)₃—N(CH₃)₂; —CH₂—N[(CH₂)₃—N(CH₃)₂]₂;—CH₂—NH—(CH₂)₃-(imidazol-1-yl); —CH₂—NH—(CH₂)₃-(morpholin-4-yl);—CH₂—NH—(CH₂)₄—NHC(NH)NH₂; —CH₂—N-(2-amino-2-deoxygluconic acid);—CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; —CH₂—NH—CH(COOH)CH₂COOH;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₈CH₃;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₉CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃;—CH₂—NH—CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—O—CH₂CH₂OH;—CH₂—NH—CH₂CH₂C(O)—N-(D-glucosamine);—CH₂—NH-(6-oxo-[1,3]oxazinan-3-yl); —CH₂—NH—CH₂CH₂—S—(CH₂)₇CH₃;—CH₂—NH—CH₂CH₂—S—(CH₂)₈CH₃; —CH₂—NH—CH₂CH₂—S—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—S—(CH₂)₁₁CH₃; —CH₂—NH—CH₂CH₂—S—(CH₂)₆Ph;—CH₂—NH—CH₂CH₂—S—(CH₂)₈Ph; —CH₂—NH—CH₂CH₂—S—(CH₂)₁₀Ph;—CH₂—NH—CH₂CH₂—S—CH₂-(4-(4-CF₃-Ph)Ph); —CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; or—CH₂—NH—(CH₂)₅—COOH. More preferably, R⁵ is hydrogen.

Preferably, R⁸ is —CH₂C(O)NH₂, —CH₂COOH, benzyl, 4-hydroxyphenyl or3-choloro-4-hydroxyphenyl.

Preferably, R⁹ is hydrogen or alkyl.

Preferably, R¹⁰ is alkyl or substituted alkyl. More preferably, R¹⁰ isthe side-chain of a naturally occurring amino acid, such as isobutyl.

Preferably, R¹¹ is hydrogen or alkyl.

Preferably, R¹² is hydrogen, alkyl, substituted alkyl or —C(O)R^(d). R¹²can also preferably be hydrogen; —CH₂COOH; —CH₂—[CH(OH)]₅CH₂OH;—CH₂CH(OH)CH₂OH; —CH₂CH₂NH₂; —CH₂C(O)OCH₂CH₃; —CH₂-(2-pyridyl);—CH₂—[CH(OH)]₄COOH; —CH₂-(3-carboxyphenyl); (R)—C(O)CH(NH₂)(CH₂)₄NH₂;—C(O)Ph; —C(O)CH₂NHC(O)CH₃; E-CH₂CH₂—S—(CH₂)₃CH═CH(CH₂)₄CH₃; or—C(O)CH₃.

Preferably, X¹ and X² are each chloro.

Preferably, X³ is hydrogen.

Preferably, each Y is independently selected from the group consistingof oxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)C(O)—, —OSO₂—,—OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂O—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)C(O)NR^(c)—,—OC(O)NR^(c)—, and —NR^(c)SO₂NR^(c)—. More preferably, Y is oxygen,sulfur or —NR^(c)—.

Preferably, n is 0 or 1, and more preferably, n is 1.

Another preferred group of glycopeptide antibiotics for use in thisinvention are those of formula II:

wherein:

R¹⁹ is hydrogen;

R²⁰ is —R^(a)—Y—R^(b)-(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)-(Z)_(x); and

R^(a), Y, R^(b), Z, x, R^(f), R³, and R⁵ have any of the values orpreferred values described herein;

or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

Preferably, R²⁰ is —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃ (trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃;—CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph; —CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph; —CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph;—CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph; or —CH₂-4-(4-Cl-Ph)-Ph. Preferably,R²⁰ can also be 4-(4-chlorophenyl)benzyl or 4-(4-chlorobenzyloxy)benzyl.

Still another preferred group of glycopeptide antibiotics for use inthis invention are derivatives of the glycopeptide antibiotic A82846B(also known as chloroorienticin A oy LY264826). See for example R.Nagarajan et al., J. Org. Chem., 1988, 54, 983–986; and N. Tsuji et al.,J. Antibiot., 1988, 41, 819–822. The structure of this glycopeptide issimilar to vancomycin, except A82846B contains an additional amino sugar(i.e. 4-epi-vancosamine attached at the R² position in formula I.) andfurther contains 4-epi-vancosamine in place of vancosamine in thedisaccharide moiety attached at the R¹ position in formula I. Forexample, a preferred group of compounds are N-alkylated derivatives ofA82846B; or a pharmaceutically acceptable salt thereof. For example, apreferred compound is a derivative of A82846B having a4-(4-chlorophenyl)benzyl group or a 4-(4-chlorobenzyloxy)benzyl groupattached at the amino group of the 4-epi-vancosamine of the disaccharidemoiety.

The pharmaceutical compositions of this invention are highly effectivefor treating bacterial diseases. Accordingly, in one of its methodaspects, this invention is also directed to a method of treating abacterial disease in a mammal, the method comprising administering tothe mammal a pharmaceutical composition comprising cyclodextrin and atherapeutically effective amount of a glycopeptide antibiotic, or apharmaceutically acceptable salt thereof. This method includes each ofthe preferred embodiments for the pharmaceutical composition describedherein.

In another of its method aspects, this invention is directed to a methodfor reducing tissue accumulation of a glycopeptide antibiotic whenadministered to a mammal, the method comprising administering theglycopeptide antibiotic to the mammal in a pharmaceutical compositioncomprising a cyclodextrin and a therapeutically effective amount of theglycopeptide antibiotic, or a pharmaceutically acceptable salt thereof.

This invention is also directed to a method for reducing nephrotoxicityproduced by a glycopeptide antibiotic when administered to a mammal, themethod comprising administering the glycopeptide antibiotic to themammal in a pharmaceutical composition comprising a cyclodextrin and atherapeutically effective amount of the glycopeptide antibiotic, or apharmaceutically acceptable salt thereof.

This invention is also directed to a method for reducing histaminerelease produced by a glycopeptide antibiotic when administered to amammal, the method comprising administering the glycopeptide antibioticto the mammal in a pharmaceutical composition comprising a cyclodextrinand a therapeutically effective amount of the glycopeptide antibiotic,or a pharmaceutically acceptable salt thereof.

This invention is also directed to a method for reducing vascularirritation produced by a glycopeptide antibiotic when administered to amammal, the method comprising administering the glycopeptide antibioticto the mammal in a pharmaceutical composition comprising a cyclodextrinand a therapeutically effective amount of the glycopeptide antibiotic,or a pharmaceutically acceptable salt thereof.

According to one embodiment, the above described methods of theinvention can be carried out by administering the glycopeptideantibiotic to the mammal in a pharmaceutical composition comprising anaqueous cyclodextrin solution and a therapeutically effective amount ofthe glycopeptide antibiotic.

Preferably, according to the methods of the invention, the weight ratioof cyclodextrin to glycopeptide will range from about 0.5:1 to 20:1, andmore preferably, from about 1:1 to about 10:1.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to novel pharmaceutical compositions and tomethods of treating bacterial diseases in a mammal using suchcompositions. When describing the compounds, compositions and methods ofthis invention, the following terms have the following meanings, unlessotherwise indicated.

Definitions

The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain preferably having from 1 to 40 carbon atoms,more preferably 1 to 10 carbon atoms, and even more preferably 1 to 6carbon atoms. This term is exemplified by groups such as methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, n-hexyl, n-decyl, tetradecyl,and the like.

The term “substituted alkyl” refers to an alkyl group as defined above,having from 1 to 8 substituents, preferably 1 to 5 substituents, andmore preferably 1 to 3 substituents, selected from the group consistingof alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxy, carboxyalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₃H, guanido, and —SO₂-heteroaryl.

The term “alkylene” refers to a diradical of a branched or unbranchedsaturated hydrocarbon chain, preferably having from 1 to 40 carbonatoms, preferably 1–10 carbon atoms, more preferably 1–6 carbon atoms.This term is exemplified by groups such as methylene (—CH₂—), ethylene(—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₃)CH₂—)and the like.

The term “substituted alkylene” refers to an alkylene group, as definedabove, having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO₂-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkylene groupsinclude those where 2 substituents on the alkylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkylene group. Preferably such fused groups contain from 1 to 3fused ring structures. Additionally, the term substituted alkyleneincludes alkylene groups in which from 1 to 5 of the alkylene carbonatoms are replaced with oxygen, sulfur or —NR— where R is hydrogen oralkyl. Examples of substituted alkylenes are chloromethylene (—CH(Cl)—),aminoethylene (—CH(NH₂)CH₂—), 2-carboxypropylene isomers(—CH₂CH(CO₂H)CH₂—), ethoxyethyl (—CH₂CH₂O—CH₂CH₂—) and the like.

The term “alkaryl” refers to the groups-alkylene-aryl and -substitutedalkylene-aryl where alkylene, substituted alkylene and aryl are definedherein. Such alkaryl groups are exemplified by benzyl, phenethyl and thelike.

The term “alkoxy” refers to the groups alkyl-O—, alkenyl-O—,cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, where alkyl, alkenyl,cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferredalkoxy groups are alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

The term “substituted alkoxy” refers to the groups substituted alkyl-O—,substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

The term “alkylalkoxy” refers to the groups-alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylalkoxy groups are alkylene-O-alkyl and include, by way ofexample, methylenemethoxy (—CH₂OCH₃), ethylenemethoxy (—CH₂CH₂OCH₃),n-propylene-iso-propoxy (—CH₂CH₂CH₂OCH(CH₃)₂), methylene-t-butoxy(—CH₂—O—C(CH₃)₃) and the like.

The term “alkylthioalkoxy” refers to the group-alkylene-S-alkyl,alkylene-S-substituted alkyl, substituted alkylene-S-alkyl andsubstituted alkylene-S-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylthioalkoxy groups are alkylene-S-alkyl and include, byway of example, methylenethiomethoxy (—CH₂SCH₃), ethylenethiomethoxy(—CH₂CH₂SCH₃), n-propylene-iso-thiopropoxy (—CH₂CH₂CH₂SCH(CH₃)₂),methylene-t-thiobutoxy (—CH₂SC(CH₃)₃) and the like.

The term “alkenyl” refers to a monoradical of a branched or unbranchedunsaturated hydrocarbon group preferably having from 2 to 40 carbonatoms, more preferably 2 to 10 carbon atoms and even more preferably 2to 6 carbon atoms and having at least 1 and preferably from 1–6 sites ofvinyl unsaturation. Preferred alkenyl groups include ethenyl (—CH═CH₂),n-propenyl (—CH₂CH═CH₂), iso-propenyl (—C(CH₃)═CH₂), and the like.

The term “substituted alkenyl” refers to an alkenyl group as definedabove having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

The term “alkenylene” refers to a diradical of a branched or unbranchedunsaturated hydrocarbon group preferably having from 2 to 40 carbonatoms, more preferably 2 to 10 carbon atoms and even more preferably 2to 6 carbon atoms and having at least 1 and preferably from 1–6 sites ofvinyl unsaturation. This term is exemplified by groups such asethenylene (—CH═CH—), the propenylene isomers (e.g., —CH₂CH═CH— and—C(CH₃)═CH—) and the like.

The term “substituted alkenylene” refers to an alkenylene group asdefined above having from 1 to 5 substituents, and preferably from 1 to3 substituents, selected from the group consisting of alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substitutedamino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen,hydroxyl, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkenylene groupsinclude those where 2 substituents on the alkenylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkenylene group.

The term “alkynyl” refers to a monoradical of an unsaturated hydrocarbonpreferably having from 2 to 40 carbon atoms, more preferably 2 to 20carbon atoms and even more preferably 2 to 6 carbon atoms and having atleast 1 and preferably from 1–6 sites of acetylene (triple bond)unsaturation. Preferred alkynyl groups include ethynyl (—C≡CH),propargyl (—CH₂C≡CH) and the like.

The term “substituted alkynyl” refers to an alkynyl group as definedabove having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

The term “alkynylene” refers to a diradical of an unsaturatedhydrocarbon preferably having from 2 to 40 carbon atoms, more preferably2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms andhaving at least 1 and preferably from 1–6 sites of acetylene (triplebond) unsaturation. Preferred alkynylene groups include ethynylene(—C≡C—), propargylene (—CH₂C≡C—) and the like.

The term “substituted alkynylene” refers to an alkynylene group asdefined above having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl

The term “acyl” refers to the groups HC(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, cycloalkyl-C(O)—, substituted cycloalkyl-C(O)—,cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—, aryl-C(O)—,heteroaryl-C(O)— and heterocyclic-C(O)— where alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “acylamino” or “aminocarbonyl” refers to the group —C(O)NRRwhere each R is independently hydrogen, alkyl, substituted alkyl, aryl,heteroaryl, heterocyclic or where both R groups are joined to form aheterocyclic group (e.g., morpholino) wherein alkyl, substituted alkyl,aryl, heteroaryl and heterocyclic are as defined herein.

The term “aminoacyl” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

The term “aminoacyloxy” or “alkoxycarbonylamino” refers to the group—NRC(O)OR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substitutedalkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclic-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclic are as defined herein.

The term “aryl” refers to an unsaturated aromatic carbocyclic group offrom 6 to 20 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed (fused) rings, wherein at least one ring is aromatic(e.g., naphthyl, dihydrophenanthrenyl, fluorenyl, or anthryl). Preferredaryls include phenyl, naphthyl and the like.

Unless otherwise constrained by the definition for the aryl substituent,such aryl groups can optionally be substituted with from 1 to 5substituents, preferably 1 to 3 substituents, selected from the groupconsisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substitutedalkoxy, substituted alkenyl, substituted alkynyl, substitutedcycloalkyl, substituted cycloalkenyl, amino, substituted amino,aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, sulfonamide,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy.

The term “aryloxy” refers to the group aryl-O— wherein the aryl group isas defined above including optionally substituted aryl groups as alsodefined above.

The term “arylene” refers to the diradical derived from aryl (includingsubstituted aryl) as defined above and is exemplified by 1,2-phenylene,1,3-phenylene, 1,4-phenylene, 1,2-naphthylene and the like.

The term “amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl and heterocyclic provided thatboth R's are not hydrogen.

“Amino acid” refers to any of the naturally occurring amino acids (e.g.Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Hyl, Hyp, Ile, Leu, Lys,Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D, L, or DL form. Theside chains of naturally occurring amino acids are well known in the artand include, for example, hydrogen (e.g., as in glycine), alkyl (e.g.,as in alanine, valine, leucine, isoleucine, proline), substituted alkyl(e.g., as in threonine, serine, methionine, cysteine, aspartic acid,asparagine, glutamic acid, glutamine, arginine, and lysine), alkaryl(e.g., as in phenylalanine and tryptophan), substituted arylalkyl (e.g.,as in tyrosine), and heteroarylalkyl (e.g., as in histidine).

The term “carboxy” refers to —COOH.

The term “C-terminus” as it relates to a glycopeptide is well understoodin the art. For example, for a glycopeptide of formula I, the C-terminusis the position substituted by the group R³.

The term “dicarboxy-substituted alkyl” refers to an alkyl groupsubstituted with two carboxy groups. This term includes, by way ofexample, —CH₂(COOH)CH₂COOH and —CH₂(COOH)CH₂CH₂COOH.

The term “carboxyalkyl” or “alkoxycarbonyl” refers to the groups“—C(O)O-alkyl”, “—C(O)O-substituted alkyl”, “—C(O)O-cycloalkyl”,“—C(O)O-substituted cycloalkyl”, “—C(O)O-alkenyl”, “—C(O)O-substitutedalkenyl”, “—C(O)O-alkynyl” and “—C(O)O-substituted alkynyl” where alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl and substituted alkynyl alkynyl are asdefined herein.

The term “cycloalkyl” refers to cyclic alkyl groups of from 3 to 20carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups havingfrom 1 to 5 substituents, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

The term “cycloalkenyl” refers to cyclic alkenyl groups of from 4 to 20carbon atoms having a single cyclic ring and at least one point ofinternal unsaturation. Examples of suitable cycloalkenyl groups include,for instance, cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-enyl and thelike.

The term “substituted cycloalkenyl” refers to cycloalkenyl groups havingfrom 1 to 5 substituents, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

The term “halo” or “halogen” refers to fluoro, chloro, bromo and iodo.

“Haloalkyl” refers to alkyl as defined herein substituted by 1–4 halogroups as defined herein, which may be the same or different.Representative haloalkyl groups include, by way of example,trifluoromethyl, 3-fluorododecyl, 12,12,12-trifluorododecyl,2-bromooctyl, 3-bromo-6-chloroheptyl, and the like.

The term “heteroaryl” refers to an aromatic group of from 1 to 15 carbonatoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfurwithin at least one ring (if there is more than one ring).

Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 5 substituents, preferably 1 to 3 substituents, selected from thegroup consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, -substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy. Such heteroaryl groups can have a singlering (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl,pyrrolyl and furyl.

“Heteroarylalkyl” refers to (heteroaryl)alkyl- where heteroaryl andalkyl are as defined herein. Representative examples include2-pyridylmethyl and the like.

The term “heteroaryloxy” refers to the group heteroaryl-O—.

The term “heteroarylene” refers to the diradical group derived fromheteroaryl (including substituted heteroaryl), as defined above, and isexemplified by the groups 2,6-pyridylene, 2,4-pyridiylene,1,2-quinolinylene, 1,8-quinolinylene, 1,4-benzofuranylene,2,5-pyridnylene, 2,5-indolenyl and the like.

The term “heterocycle” or “heterocyclic” refers to a monoradicalsaturated or unsaturated group having a single ring or multiplecondensed rings, from 1 to 40 carbon atoms and from 1 to 10 heteroatoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 5, and preferably 1 to 3 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, oxo (═O), and—SO₂-heteroaryl. Such heterocyclic groups can have a single ring ormultiple condensed rings. Preferred heterocyclics include morpholino,piperidinyl, and the like.

Examples of nitrogen heterocycles and heteroaryls include, but are notlimited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, indoline, morpholino, piperidinyl,tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containingheterocycles.

Another class of heterocyclics is known as “crown compounds” whichrefers to a specific class of heterocyclic compounds having one or morerepeating units of the formula [—(CH₂—)_(a)A-] where a is equal to orgreater than 2, and A at each separate occurrence can be O, N, S or P.Examples of crown compounds include, by way of example only,[—(CH₂)₃—NH—]₃, [—((CH₂)₂—O)₄—((CH₂)₂—NH)₂] and the like. Typically suchcrown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbonatoms.

The term “heterocyclooxy” refers to the group heterocyclic-O—.

The term “thioheterocyclooxy” refers to the group heterocyclic-S—.

The term “oxyacylamino” or “aminocarbonyloxy” refers to the group—OC(O)NRR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substitutedalkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “phosphono” refers to —PO₃H₂.

The term “prodrug” is well understood in the art and includes compoundsthat are converted to pharmaceutically active compounds of the inventionin a mammalian system. For example, see Remington's PharmaceuticalSciences, 1980, vol. 16, Mack Publishing Company, Easton, Pa., 61 and424.

The term “saccharide group” refers to an oxidized, reduced orsubstituted saccharide monoradical covalently attached to theglycopeptide or other compound via any atom of the saccharide moiety,preferably via the aglycone carbon atom. The term includesamino-containing saccharide groups. Representative saccharides include,by way of illustration, hexoses such as D-glucose, D-mannose, D-xylose,D-galactose, vancosamine, 3-desmethyl-vancosamine, 3-epi-vancosamine,4-epi-vancosamine, acosamine, actinosamine, daunosamine,3-epi-daunosamine, ristosamine, D-glucamine, N-methyl-D-glucamine,D-glucuronic acid, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine,sialyic acid, iduronic acid, L-fucose, and the like; pentoses such asD-ribose or D-arabinose; ketoses such as D-ribulose or D-fructose;disaccharides such as 2-O-(α-L-vancosaminyl)-β-D-glucopyranose,2-O-(3-desmethyl-α-L-vancosaminyl)-β-D-glucopyranose, sucrose, lactose,or maltose; derivatives such as acetals, amines, acylated, sulfated andphosphorylated sugars; oligosaccharides having from 2 to 10 saccharideunits. For the purposes of this definition, these saccharides arereferenced using conventional three letter nomenclature and thesaccharides can be either in their open or preferably in their pyranoseform.

The term “amino-containing saccharide group” or “amino saccharide”refers to a saccharide group having an amino substituent. Representativeamino-containing saccharides include L-vancosamine,3-desmethyl-vancosamine, 3-epi-vancosamine, 4-epi-vancosamine,acosamine, actinosamine, daunosamine, 3-epi-daunosamine, ristosamine,N-methyl-D-glucamine and the like.

The term “spiro-attached cycloalkyl group” refers to a cycloalkyl groupattached to another ring via one carbon atom common to both rings.

The term “stereoisomer” as it relates to a given compound is wellunderstood in the art, and refers another compound having the samemolecular formula, wherein the atoms making up the other compound differin the way they are oriented in space, but wherein the atoms in theother compound are like the atoms in the given compound with respect towhich atoms are joined to which other atoms (e.g. an enantiomer, adiastereomer, or a geometric isomer). See for example, Morrison andBoyde Organic Chemistry, 1983, 4th ed, Allyn and Bacon, Inc., Boston,Mass., page 123.

The term “sulfonamide” refers to a group of the formula —SO₂NRR, whereeach R is independently hydrogen, alkyl, substituted alkyl, aryl,heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

The term “thiol” refers to the group —SH.

The term “thioalkoxy” refers to the group —S-alkyl.

The term “substituted thioalkoxy” refers to the group —S-substitutedalkyl.

The term “thioaryloxy” refers to the group aryl-S— wherein the arylgroup is as defined above including optionally substituted aryl groupsalso defined above.

The term “thioheteroaryloxy” refers to the group heteroaryl-S— whereinthe heteroaryl group is as defined above including optionallysubstituted aryl groups as also defined above.

The term “thioether derivatives” when used to refer to the glycopeptidecompounds of this invention includes thioethers (—S—), sulfoxides (—SO—)and sulfones (—SO₂—).

As to any of the above groups which contain one or more substituents, itis understood, of course, that such groups do not contain anysubstitution or substitution patterns which are sterically impracticaland/or synthetically non-feasible. In addition, the compounds of thisinvention include all stereochemical isomers arising from thesubstitution of these compounds.

“Cyclodextrin” refers to cyclic molecules containing six or moreα-D-glucopyranose units linked at the 1,4 positions by α linkages as inamylose. β-Cyclodextrin or cycloheptaamylose contains sevenα-D-glucopyranose units. As used herein, the term “cyclodextrin” alsoincludes cyclodextrin derivatives such as hydroxypropyl and sulfobutylether cyclodextrins, and others. Such derivatives are described forexample, in U.S. Pat. Nos. 4,727,064 and 5,376,645. Additionally,hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin arecommercially available. One preferred cyclodextrin is hydroxypropylβ-cyclodextrin having a degree of substitution of from about 4.1–5.1 asmeasured by FTIR. Such a cyclodextrin is available from Cerestar(Hammond, Ind., USA) under the name Cavitron™ 82003.

The term “aqueous cyclodextrin carrier” refers to an aqueouscyclodextrin solution comprising a cyclodextrin and water.

“Glycopeptide” refers to oligopeptide (e.g. heptapeptide) antibiotics,characterized by a multi-ring peptide core optionally substituted withsaccharide groups, such as vancomycin. Examples of glycopeptidesincluded in this definition may be found in “GlycopeptidesClassification, Occurrence, and Discovery”, by Raymond C. Rao and LouiseW. Crandall, (“Drugs and the Pharmaceutical Sciences” Volume 63, editedby Ramakrishnan Nagarajan, published by Marcal Dekker, Inc.). Additionalexamples of glycopeptides are disclosed in U.S. Pat. Nos. 4,639,433;4,643,987; 4,497,802; 4,698,327; 5,591,714; 5,840,684; and 5,843,889; inEP 0 802 199; EP 0 801 075; EP 0 667 353; WO 97/28812; WO 97/38702; WO98/52589; WO 98/52592; and in J. Amer. Chem. Soc., 1996, 118,13107–13108; J. Amer. Chem. Soc., 1997, 119, 12041–12047; and J. Amer.Chem. Soc., 1994, 116, 4573–4590. Representative glycopeptides includethose identified as A477, A35512, A40926, A41030, A42867, A47934,A80407, A82846, A83850, A84575, AB-65, Actaplanin, Actinoidin, Ardacin,Avoparcin, Azureomycin, Balhimycin, Chloroorientiein, Chloropolysporin,Decaplanin, N-demethylvancomycin, Eremomycin, Galacardin, Helvecardin,Izupeptin, Kibdelin, LL-AM374, Mannopeptin, MM45289, MM47756, MM47761,MM49721, MM47766, MM55260, MM55266, MM55270, MM56597, MM56598, OA-7653,Orenticin, Parvodicin, Ristocetin, Ristomycin, Synmonicin, Teicoplanin,UK-68597, UK-69542, UK-7205 1, Vancomycin, and the like. The term“glycopeptide” or “glycopeptide antibiotic” as used herein is alsointended to include the general class of glycopeptides disclosed aboveon which the sugar moiety is absent, i.e. the aglycone series ofglycopeptides. For example, removal of the disaccharide moiety appendedto the phenol on vancomycin by mild hydrolysis gives vancomycinaglycone. Also included within the scope of the term “glycopeptideantibiotics” are synthetic derivatives of the general class ofglycopeptides disclosed above, included alkylated and acylatedderivatives. Additionally, within the scope of this term areglycopeptides that have been further appended with additional saccharideresidues, especially aminoglycosides, in a manner similar tovancosamine.

The term “lipidated glycopeptide” refers specifically to thoseglycopeptide antibiotics which have been synthetically modified tocontain a lipid substituent. As used herein, the term “lipidsubstituent” refers to any substituent contains 5 or more carbon atoms,preferably, 10 to 40 carbon atoms. The lipid substituent may optionallycontain from 1 to 6 heteroatoms selected from halo, oxygen, nitrogen,sulfur and phosphorous. Lipidated glycopeptide antibiotics arewell-known in the art. See, for example, in U.S. Pat. Nos. 5,840,684,5,843,889, 5,916,873, 5,919,756, 5,952,310, 5,977,062, 5,977,063, and EP667, 353, WO 98/52589, WO 99/56760, WO 00/04044, WO 00/39156, thedisclosures of which are incorporated herein by reference in theirentirety.

“Vancomycin” refers to a glycopeptide antibiotic having the formula:

When describing vancomycin derivatives, the term “N^(van)-” indicatesthat a substituent is covalently attached to the amino group of thevacosamine moiety of vacomycin. Similarly, the term “N^(leu)-” indicatesthat a substituent is covalently attached to the amino group of theleucine moiety of vancomycin.

“Optional” or “optionally” means that the subsequently described eventor circumstance may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, “optionally substituted” means that a groupmay or may not be substituted with the described substituent.

As used herein, the terms “inert organic solvent” or “inert solvent” or“inert diluent” mean a solvent or diluent which is essentially inertunder the conditions of the reaction in which it is employed as asolvent or diluent. Representative examples of materials which may beused as inert solvents or diluents include, by way of illustration,benzene, toluene, acetonitrile, tetrahydrofuran (“THF”),dimethylformamide (“DMF”), chloroform (“CHCl₃”), methylene chloride (ordichloromethane or “CH₂Cl₂), diethyl ether, ethyl acetate, acetone,methylethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

The term “nitrogen-linked” or “N-linked” means a group or substituent isattached to the remainder of a compound (e.g. a compound of formula I)through a bond to a nitrogen of the group or substituent. The term“oxygen-linked” means a group or substituent is attached to theremainder of a compound (e.g. a compound of formula I) through a bond toan oxygen of the group or substituent. The term “sulfur-linked” means agroup or substituent is attached to the remainder of a compound (e.g. acompound of formula I) through a bond to a sulfur of the group orsubstituent. “Pharmaceutically acceptable salt” means those salts whichretain the biological effectiveness and properties of the parentcompounds and which are not biologically or otherwise harmful as thedosage administered. The compounds of this invention are capable offorming both acid and base salts by virtue of the presence of amino andcarboxy groups, respectively.

Pharmaceutically acceptable base addition salts may be prepared frominorganic and organic bases. Salts derived from inorganic bases include,but are not limited to, the sodium, potassium, lithium, ammonium,calcium, and magnesium salts. Salts derived from organic bases include,but are not limited to, salts of primary, secondary and tertiary amines,substituted amines including naturally-occurring substituted amines, andcyclic amines, including isopropylamine, trimethyl amine, diethylamine,triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol,tromethamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,N-alkylglucamines, theobromine, purines, piperazine, piperidine, andN-ethylpiperidine. It should also be understood that other carboxylicacid derivatives would be useful in the practice of this invention, forexample carboxylic acid amides, including carboxamides, lower alkylcarboxamides, di(lower alkyl) carboxamides, and the like.

Pharmaceutically acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid and the like.

The compounds of this invention typically contain one or more chiralcenters. Accordingly, this invention is intended to include racemicmixtures, diasteromers, enantiomers and mixture enriched in one or moresteroisomer. The scope of the invention as described and claimedencompasses the racemic forms of the compounds as well as the individualenantiomers and non-racemic mixtures thereof.

The term “treatment” as used herein includes any treatment of acondition or disease in an animal, particularly a mammal, moreparticularly a human, and includes:

(i) preventing the disease or condition from occurring in a subjectwhich may be predisposed to the disease but has not yet been diagnosedas having it;

(ii) inhibiting the disease or condition, i.e. arresting itsdevelopment; relieving the disease or condition, i.e. causing regressionof the condition; or relieving the conditions caused by the disease,i.e. symptoms of the disease.

The term “disease state which is alleviated by treatment with a broadspectrum antibacterial” or “bacterial disease” as used herein isintended to cover all disease states which are generally acknowledged inthe art to be usefully treated with a broad spectrum antibacterial ingeneral, and those disease states which have been found to be usefullytreated by the specific antibacterials of this invention. Such diseasestates include, but are not limited to, treatment of a mammal afflictedwith pathogenic bacteria, in particular staphylococci (methicillinsensitive and resistant), streptococci (penicillin sensitive andresistant), enterococci (vancomycin sensitive and resistant), andClostridium difficile

The term “therapeutically effective amount” refers to that amount whichis sufficient to effect treatment, as defined herein, when administeredto a mammal in need of such treatment. The therapeutically effectiveamount will vary depending on the subject and disease state beingtreated, the severity of the affliction and the manner ofadministration, and may be determined routinely by one of ordinary skillin the art.

The term “protecting group” or “blocking group” refers to any groupwhich, when bound to one or more hydroxyl, thiol, amino, carboxy orother groups of the compounds, prevents undesired reactions fromoccurring at these groups and which protecting group can be removed byconventional chemical or enzymatic steps to reestablish the hydroxyl,thio, amino, carboxy or other group. The particular removable blockinggroup employed is not critical and preferred removable hydroxyl blockinggroups include conventional substituents such as allyl, benzyl, acetyl,chloroacetyl, thiobenzyl, benzylidine, phenacyl, t-butyl-diphenylsilyland any other group that can be introduced chemically onto a hydroxylfunctionality and later selectively removed either by chemical orenzymatic methods in mild conditions compatible with the nature of theproduct. Protecting groups are disclosed in more detail in T. W. Greeneand P. G. M. Wuts, “Protective Groups in Organic Synthesis” 3^(rd) Ed.,1999, John Wiley and Sons, N.Y.

Preferred removable amino blocking groups include conventionalsubstituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ),fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like,which can be removed by conventional conditions compatible with thenature of the product.

Preferred carboxy protecting groups include esters such as methyl,ethyl, propyl, t-butyl etc. which can be removed by mild conditionscompatible with the nature of the product.

General Synthetic Procedures

The glycopeptide antibiotics employed in this invention are commerciallyavailable or can be prepared from readily available starting materialsusing the following general methods and procedures. It will beappreciated that where typical or preferred process conditions (i.e.,reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given, other process conditions can also be usedunless otherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and G. M. Wuts, ProtectingGroups in Organic Synthesis, Third Edition, Wiley, New York, 1999, andreferences cited therein.

In the following reaction schemes, the glycopeptide compounds aredepicted in a simplified form as a box “G” that shows the carboxyterminus labeled [C], the vancosamine amino terminus labeled [V], the“non-saccharide” amino terminus (leucine amine moiety) labeled [N], andoptionally, the resorcinol moiety labeled [R] as follows:

By way of illustration, a lipidated glycopeptide compound useful in thepresent invention can be prepared by reductive alkylated as shown in thefollowing reaction:

where A represents R^(a) minus one carbon atom and R^(a), R^(b), Y, Zand x are as defined herein. This reaction is typically conducted byfirst contacting one equivalent of the glycopeptide, i.e., vancomycin,with an excess, preferably from 1.1 to 1.3 equivalents, of the desiredaldehyde in the presence of an excess, preferably about 2.0 equivalents,of a tertiary amine, such as diisopropylethylamine (DIPEA) and the like.This reaction is typically conducted in an inert diluent, such as DMF oracetonitrile/water, at ambient temperature for about 0.25 to 2 hoursuntil formation of the corresponding imine and/or hemiaminal issubstantially complete. The resulting imine and/or hemiaminal istypically not isolated, but is reacted in situ with a metal hydridereducing agent, such as sodium cyanoborohydride and the like, to affordthe corresponding amine. This reaction is preferably conducted bycontacting the imine and/or hemiaminal with an excess, preferably about3 equivalents, of trifluoroacetic acid, followed by about 1 to 1.2equivalents of the reducing agent at ambient temperature in methanol oracetonitrile/water. The resulting alkylated product is readily purifiedby conventional procedures, such as precipitation and/or reverse-phasehigh-performance liquid chromatography (HPLC). Surprisingly, by formingthe imine and/or hemiaminal in the presence of a trialkyl amine, andthen acidifying with trifluoroacetic acid before contact with thereducing agent, the selectivity for the reductive alkylation reaction isgreatly improved, i.e., reductive alkylation at the amino group of thesaccharide (e.g., vancosamine) is favored over reductive alkylation atthe N-terminus (e.g., the leucinyl group) by at least 10:1, morepreferably 20:1.

The reductive alkylation process is typically carried out in thepresence of a suitable solvent or combination of solvents, such as, forexample, a halogenated hydrocarbon (e.g. methylene chloride), a linearor branched ether (e.g. diethyl ether, tetrahydrofuran), an aromatichydrocarbon (e.g. benzene or toluene), an alcohol (methanol, ethanol, orisopropanol), dimethylsulfoxide (DMSO), N,N-dimethylformamide,acetonitrile, water, 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidone,tetramethyl urea, N,N-dimethylacetamide, diethylformamide (DMF),1-methyl-2-pyrrolidinone, tetramethylenesulfoxide, glycerol, ethylacetate, isopropyl acetate, N,N-dimethylpropylene urea (DMPU) ordioxane. Preferably the alkylation is carried out in acetonitrile/water,or DMF/methanol.

Preferably the reduction (i.e. treatment with the reducing agent) iscarried out in the presence of a protic solvent, such as, for example,an alcohol (e.g. methanol, ethanol, propanol, isopropanol, or butanol),water, or the like.

The reductive alkylation process of the invention can be carried out atany suitable temperature from the freezing point to the refluxtemperature of the reaction mixture. Preferably the reaction is carriedout at a temperature in the range of about 0° C. to about 100° C. Morepreferably at a temperature in a range of about 0° C. to about 50° C.,or in a range of about 20° C. to about 30° C.

Any suitable base can be employed in the reductive alkylation process ofthe invention. Suitable bases include tertiary amines (e.g.diisopropylethylamine, N-methylmorpholine or triethylamine) and thelike.

Any suitable acid can be used to acidify the reaction mixture. Suitableacids include carboxylic acids (e.g. acetic acid, trichloroacetic acid,citric acid, formic acid, or trifluoroacetic acid), mineral acids (e.g.hydrochloric acid, sulfuric acid, or phosphoric acid), and the like. Apreferred acid is trifluoroacetic acid.

Suitable reducing agents for carrying out reductive alkylation processof the invention are known in the art. Any suitable reducing agent canbe employed in the methods of the invention, provided it is compatiblewith the functionality present in the glycopeptide. For example,suitable reducing agents include sodium cyanoborohydride,triacetoxyborohydride, pyridine/borane, sodium borohydride, and zincborohydride. The reduction can also be carried out in the presence of atransition metal catalyst (e.g. palladium or platinum) in the presenceof a hydrogen source (e.g. hydrogen gas or cycloheadiene). See forexample, Advanced Organic Chemistry, Fourth Edition, John Wiley & Sons,New York (1992), 899–900.

Any glycopeptide having an amino group may be employed in thesereductive alkylation reactions. Such glycopeptides are well-known in theart and are either commercially available or may be isolated usingconventional procedures. Suitable glycopeptides are disclosed, by way ofexample, in U.S. Pat. Nos. 3,067,099; 3,338,786; 3,803,306; 3,928,571;3,952,095; 4,029,769; 4,051,237; 4,064,233; 4,122,168; 4,239,751;4,303,646; 4,322,343; 4,378,348; 4,497,802; 4,504,467; 4,542,018;4,547,488; 4,548,925; 4,548,974; 4,552,701; 4,558,008; 4,639,433;4,643,987; 4,661,470; 4,694,069; 4,698,327; 4,782,042; 4,914,187;4,935,238; 4,946,941; 4,994,555; 4,996,148; 5,187,082; 5,192,742;5,312,738; 5,451,570; 5,591,714; 5,721,208; 5,750,509; 5,840,684; and5,843,889. Preferably, the glycopeptide employed in the above reactionis vancomycin.

The aldehydes and ketones employed in the above reactive alkylationreactions are also well-known in the art and are either commerciallyavailable or can be prepared by conventional procedures usingcommercially available starting materials and conventional reagents (forexample see March, Advanced Organic Chemistry, Fourth Edition, JohnWiley & Sons, New York (1992), and references cited therein). Aldehydesand ketones other than those shown above may be employed in thisalkylation reaction, including by way of example, aldehydes of theformula HC(O)—R^(f), where R^(f) is as defined herein.

If desired, aminoalkyl sidechain can be introduced at the resorcinolmoiety of a glycopeptide, such as vancomycin, via a Mannich reaction. Inthis reaction, an amine of formula NHRR′ (wherein one or both of R andR′ is a alkyl or substituted alkyl group or a group that comprises oneor more phosphono groups), and an aldehyde (CH₂O), such as formalin (asource of formaldehyde), are reacted with the glycopeptide under basicconditions to give the glycopeptide derivative, as shown below (in thisscheme, the resorcinol moiety is shown for clarity).

When employed, the phosphono substituted compounds (e.g. the phosphonosubstituted amines, alcohols, or thiols) are either commerciallyavailable or can be prepared by conventional procedures usingcommercially available starting materials and reagents. See for example,Advanced Organic Chemistry, Jerry March, 4th ed, 1992, John Wiley andSons, New York, page 959; and Frank R. Hartley (ed.) The Chemistry ofOrganophosphorous Compounds, vol. 1–4, John Wiley and Sons, New York(1996). Aminomethylphosphonic acid is commercially available fromAldrich Chemical Company, Milwaukee, Wis.

Additional details and other methods for preparing the compounds of thisinvention are described in the Examples below.

Pharmaceutical Compositions

The pharmaceutical compositions of this invention comprise aglycopeptide antibiotic and a cyclodextrin compound. Preferably, thepharmaceutical compositions of this invention are formulated forparenteral administration for the therapeutic or prophylactic treatmentof bacterial diseases.

By way of illustration, the glycopeptide antibiotic, preferably in theform a pharmaceutically acceptable salt, can be admixed with an aqueouscyclodextrin solution to form a composition of this invention. Suchpharmaceutical compositions will typically contain from about 1 to about40 weight percent of the cyclodextrin and a therapeutically effectiveamount of the glycopeptide antibiotic.

Optionally, the pharmaceutical composition may contain otherpharmaceutically acceptable components, such a buffers, surfactants,antioxidants, viscosity modifying agents, preservatives and the like.Each of these components is well-known in the art. See, for example,U.S. Pat. No. 5,985,310. Other components suitable for use in theformulations of the present invention can be found in Remington'sPharmaceutical Sciences, Mace Publishing Company, Philadelphia, Pa.,17th ed. (1985). In a preferred embodiment, the aqueous cyclodextrinsolution further comprises dextrose, preferably, 5% dextrose.

The glycopeptide antibiotics used in this invention are effective over awide dosage range and are typically administered in a therapeuticallyeffective amount. It, will be understood, however, that the amount ofthe compound actually administered will be determined by a physician, inthe light of the relevant circumstances, including the condition to betreated, the chosen route of administration, the actual compoundadministered and its relative activity, the age, weight, and response ofthe individual patient, the severity of the patient's symptoms, and thelike.

Suitable doses are in the general range of from 0.01–100 mg/kg/day,preferably 0.1–50 mg/kg/day. For an average 70 kg human, this wouldamount to 0.7 mg to 7 g per day, or preferably 7 mg to 3.5 g per day. Amore preferred dose for a human is about 500 mg to about 2 g per day.

The following formulation examples illustrate representativepharmaceutical compositions of the invention.

FORMULATION EXAMPLE A

An injectable preparation is prepared having the following composition:

Ingredients Glycopeptide Antibiotic 0.1–5.0 gHydroxypropyl-β-cyclodextrin 1–25 g 5% Aqueous Dextrose Solution(sterile) q.s. to 100 mL The above ingredients are blended and the pH isadjusted to 3.5 ± 0.5 using 0.5 N HCl or 0.5 N NaOH.

FORMULATION EXAMPLE B

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A frozen solution suitable for injection is prepared having thefollowing composition:

Frozen Solution Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g Excipients - e.g., dextrose 0–50 g Water for Injection 10–100 mL The weight ratio ofhydroxypropyl-β-cyclodextrin to the active compound will typically befrom about 1:1 to about 10:1.Representative Procedure: Hydroxypropyl-β-cyclodextrin and excipients,if any, are dissolved in about 80% of the water for injection and theactive compound is added and dissolved. The pH is adjusted with 1 Msodium hydroxide to 4.7±0.3 and the volume is then adjusted to 95% ofthe final volume with water for injection. The pH is checked andadjusted, if necessary, and the volume is adjusted to the final volumewith water for injection. The formulation is then sterile filteredthrough a 0.22 micron filter and placed into a sterile vial underaseptic conditions. The vial is capped, labeled and stored frozen.

FORMULATION EXAMPLE C

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A lyophilized powder useful for preparing an injectable solution isprepared having the following composition:

Lyophilized Powder Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g Excipients - e.g., mannitol,0–50 g sucrose and/or lactose Buffer agent - e.g., citrate 0–500 mg Theweight ratio of hydroxypropyl-β-cyclodextrin to the active compound willtypically be from about 1:1 to about 10:1.Representative Procedure: Hydroxypropyl-β-cyclodextrin and excipientsand/or buffering agents, if any, are dissolved in about 60% of the waterfor injection. The active compound is added and dissolved and the pH isadjusted with 1 M sodium hydroxide to 4.0–5.0 and the volume is adjustedto 95% of the final volume with water for injection. The pH is checkedand adjusted, if necessary, and the volume is adjusted to the finalvolume with water for injection. The formulation is then sterilefiltered through a 0.22 micron filter and placed into a sterile vialunder aseptic conditions. The formulation is then freeze-dried using anappropriate lyophilization cycle. The vial is capped (optionally underpartial vacuum or dry nitrogen), labeled and stored at room temperatureor under refrigeration.

FORMULATION EXAMPLE D

This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

A sterile powder useful for preparing an injectable solution is preparedhaving the following composition:

Sterile Powder Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g¹ Excipients optional Theweight ratio of hydroxypropyl-β-cyclodextrin to the active willtypically be from about 1:1 to about 10:1.Representative Procedure: Hydroxypropyl-β-cyclodextrin and the activecompound (and any excipients) are dispersed into an appropriate sterilecontainer and the container is sealed (optionally under partial vacuumor dry nitrogen), labeled and stored at room temperature or underrefrigeration.Administration of Representative Formulations C and D to a Patient

The pharmaceutical formulations described in formulation examples H andI above can be administered intravenously to a patient by theappropriate medical personnel to treat or prevent gram-positiveinfections. For administration, the above formulations can bereconstituted and/or diluted with a diluent, such as 5% dextrose orsterile saline, as follows:

Representative Procedure: The lyophilized powder of formulation exampleC (e.g., containing 1000 mg of active compound) is reconstituted with 20mL of sterile water and the resulting solution is further diluted with80 mL of sterile saline in a 100 mL infusion bag. The diluted solutionis then administered to the patient intravenously over 30 to 120minutes.Utility

The pharmaceutical compositions of this invention are useful in medicaltreatments and exhibit biological activity, including antibacterialactivity, which can be demonstrated in using the tests described herein.Such tests are well known to those skilled in the art, and arereferenced and described in Lorian “Antibiotics in Laboratory Medicine”,Fourth Edition, Williams and Wilkins (1991).

Accordingly, this invention provides methods for treating bacterial orinfectious diseases, especially those caused by Gram-positivemicroorganisms, in animals. Depending on the glycopeptide antibiotic,the compositions of this invention are particularly useful in treatinginfections caused by methicillin-resistant staphylococci.

The animal treated with the pharmaceutical compositions of thisinvention may be either susceptible to, or infected with, themicroorganism. The method of treatment typically comprises administeringto the animal a pharmaceutical composition comprising a therapeuticallyeffective amount of the glycopeptide antibiotic compound.

In practicing this method, the pharmaceutical composition can beadministered in a single daily dose or in multiple doses per day. Thetreatment regimen may require administration over extended periods oftime, for example, for several days or for from one to six weeks. Theamount per administered dose or the total amount administered willdepend on such factors as the nature and severity of the infection, theage and general health of the patient, the tolerance of the patient tothe antibiotic and the microorganism or microorganisms in the infection.

Among other properties, when administered to a mammal, thepharmaceutical compositions of this invention have been found to exhibitone or more of the following properties (a) reduced tissue accumulationof the glycopeptide antibiotic, (b) reduced nephrotoxicity, (c) reducedhistamine release (Red Man Syndrome) and (d) reduced vascularirritation, compared to a pharmaceutical composition which does notcontain the cyclodextrin. Additionally, the cyclodextrin has been foundto increase the water solubility of certain glycopeptide antibiotics,such as lipidated glycopeptide antibiotics.

The following synthetic and biological examples are offered toillustrate this invention and are not to be construed in any way aslimiting the scope of this invention.

EXAMPLES

In the examples below, the following abbreviations have the followingmeanings. Any abbreviations not defined have their generally acceptedmeaning. Unless otherwise stated, all temperatures are in degreesCelsius.

-   -   ACN=acetonitrile    -   BOC, Boc=tert-butoxycarbonyl    -   DIBAL-H=diisobutylaluminum hydride    -   DIPEA=diisopropylethylamine    -   DMF=N,N-dimethylformamide    -   DMSO=dimethyl sulfoxide    -   eq.=equivalent    -   EtOAc=ethyl acetate    -   Fmoc=9-fluorenylmethoxycarbonyl    -   HOBT=1-hydroxybenzotriazole hydrate    -   Me=methyl    -   PyBOP=benzotriazol-1-yloxytris(pyrrolidino)phosphonium        hexafluorophosphate    -   TEMPO=2,2,6,6tetramethyl-piperidinyloxy, free radical    -   TFA=trifluoroacetic acid    -   THF=tetrahydrofuran    -   TLC, tlc=thin layer chromatography

In the following examples, vancomycin hydrochloride semi-hydrate waspurchased from Alpharma, Inc. Fort Lee, N.J. 07024 (Alpharma AS, OsloNorway). Other reagents and reactants are available from AldrichChemical Co., Milwaukee, Wis. 53201.

General Procedure A Reductive Alkylation of Vancomycin

To a mixture of vancomycin (1 eq.) and the desired aldehyde (1.3 eq.) inDMF was added DIPEA (2 eq.). The reaction was stirred at ambienttemperature for 1–2 hours and monitored by reverse-phase HPLC. Methanoland NaCNBH₃ (1 eq.) were added to the solution, followed by TFA (3 eq.).Stirring was continued for an additional hour at ambient temperature.After the reaction was complete, the methanol was removed in vacuo. Theresidue was precipitated in acetonitrile. Filtration gave the crudeproduct which was then purified by reverse-phase HPLC. If desired, otherglycopeptides antibiotics may be used in this procedure.

Example 1 Preparation of Compound A (Formula II Wherein R³ is —OH; R⁵N-(phosphonomethyl)aminomethyl; R¹⁹ is Hydrogen, and R²⁰ is—CH₂CH₂—NH—(CH₂)₉CH₃)

(Aminomethyl)phosphonic acid (3.88 g, 35 mmol) and diisopropylethylamine(6.1 ml, 35 mmol) were combined in water (40 ml) and stirred untilhomogeneous. Acetonirile (50 ml) and formaldehyde (37% solution in H₂O;0.42 ml, 05.6 mmol) were then added to the reaction mixture. Afterapproximately 15 minutes both N^(VAN)-decylaminoethyl vancomycintristrifluoroacetate (10.0 g, 5.1 mmol) and diisopropylethylamine (6.1ml, 35 mmol) were added to the reaction mixture. The reaction wasstirred at room temperature for approximately 18 hrs, at which time theacetonitrile was removed in vacuo, and the residue was lyophylized. Theresulting solid was triturated with water (100 mL), collected byfiltration, dried in vacuo and purified by reverse phase preparativeHPLC to give the title compound. MS calculated (MH+) 1756.6; found (MH+)1756.6.

Example 2 Preparation of Pharmaceutical Compositions

Injectable pharmaceutical compositions can be prepared as follows:

5% Aqueous Hydroxypropyl-β- Dextrose Compound A cyclodextrin Solution(sterile) 2A 200 mg  1 g q.s. to 100 mL 2B 200 mg  5 g q.s. to 100 mL 2C200 mg 25 g q.s. to 100 mL 2D 200 mg 30 g q.s. to 100 mL

The above ingredients were blended and the pH was adjusted to 3.5±0.5using 0.5 N HCl or 0.5 N NaOH.

Example 3 Preparation of a Glycopeptide that can be Incorporated into aComposition of the Invention (Formula II Wherein R³ is —OH; R⁵ is H; R¹⁹is Hydrogen, and R²⁰ is 4-(4-chlorophenyl)benzyl

A three liter 3-necked flask was fitted with a condenser, nitrogen inletand overhead mechanical stirring apparatus. The flask was charged withpulverized A82846B acetate salt (20.0 g, 1.21×10⁻⁵ mol) and methanol(1000 mL) under a nitrogen atmosphere, 4′-chlorobiphenylcarboxaldehyde(2.88 g, 1.33×10⁻² mol, 1.1 eq.) was added to this stirred mixture,followed by methanol (500 mL). Finally, sodium cyanoborohydride (0.84 g,1.33×10⁻² mol, 1.1 eq.) was added followed by methanol (500 mL). Theresulting mixture was heated to reflux (about 65° C.).

After 1 hour at reflux, the reaction mixture attained homogeneity. After25 hours at reflux, the heat source was removed and the clear reactionmixture was measured with a pH meter (6.97 at 58.0° C.). 1N NaOH (22.8mL) was added dropwise to adjust the pH to 9.0 (at 54.7° C.). The flaskwas equipped with a distillation head and the mixture was concentratedunder partial vacuum to a weight of 322.3 grams while maintaining thepot temperature between 40°–45° C.

The distillation head was replaced with an addition funnel containing500 mL of isopropanol (IPA). The IPA was added dropwise to the roomtemperature solution over 1 hour. After approximately ⅓ of the IPA wasadded, a granular precipitate formed. The remaining IPA was added at afaster rate after precipitation had commenced. The flask was weighed andfound to hold 714.4 grams of the IPA/methanol slurry.

The flask was re-equipped with a still-head and distilled under partialvacuum to remove the remaining methanol. The resulting slurry (377.8 g)was allowed to chill in the freezer overnight. The crude product wasfiltered through a polypropylene pad and rinsed twice with 25 mL of coldIPA. After pulling dry on the funnel for 5 minutes, the material wasplaced in the vacuum oven to dry ta 40° C. A light pink solid (22.87 g(theory=22.43 g)) was recovered. HPLC analysis versus a standardindicated 68.0% weight percent of the title compound(4-[4-chlorophenyl]benzyl-A82846B] in the crude solid, which translatedinto a corrected crude yield of 69.3%.

The products of the reaction were analyzed by reverse-phase HPLCutilizing a Zorbax SB-C₁₈ column with ultra-violet light (UV; 230 nm)detection. A 20 minute gradient solvent system consisting of 95% aqueousbuffer/5% CH₃CN at time=0 minutes to 40% aqueous buffer/60% CH₃CN attime=30 minutes was used, where the aqueous buffer was TEAP (5 ml CH₃CN,3 ml phosphoric acid in 1000 ml water).

The intermediate A82846B acetate salt can be prepared as described inU.S. Pat. No. 5,840,684.

Example 4 Determination of Antibacterial Activity

A. In Vitro Determination of Antibacterial Activity

1. Determination of Minimal Inhibitory Concentrations (MICs)

This procedure may be used to assess the antibacterial properties of theglycopeptide antibiotic. Bacterial strains were obtained from eitherAmerican Type Tissue Culture Collection (ATCC), Stanford UniversityHospital (SU), Kaiser Permanente Regional Laboratory in Berkeley (KPB),Massachusetts General Hospital (MGH), the Centers for Disease Control(CDC), the San Francisco Veterans' Administration Hospital (SFVA) or theUniversity of California San Francisco Hospital (UCSF). Vancomycinresistant enterococci were phenotyped as Van A or Van B based on theirsensitivity to teicoplanin. Some vancomycin resistant enterococci thathad been genotyped as Van A, Van B, Van C1 or Van C2 were obtained fromthe Mayo Clinic.

Minimal inhibitory concentrations (MICs) were measured in amicrodilution broth procedure under NCCLS guidelines. Routinely, thecompounds were serially diluted into Mueller-Hinton broth in 96-wellmicrotiter plates. Overnight cultures of bacterial strains were dilutedbased on absorbance at 600 nm so that the final concentration in eachwell was 5×10⁵ cfu/mL. Plates were returned to a 35° C. incubator. Thefollowing day (or 24 hours in the case of Enterococci strains), MICswere determined by visual inspection of the plates. Strains routinelytested in the initial screen included methicillin-sensitiveStaphylococcus aureus (MSSA), methicillin-resistant Staphylococcusaureus, methicillin-sensitive Staphylococcus epidermidis (MSSE),methicillin-resistant Staphylococcus epidermidis (MRSE), vancomycinsensitive Enterococcus faecium (VSE Fm), vancomycin sensitiveEnterococcus faecalis (VSE Fs), vancomycin resistant Enterococcusfaecium also resistant to teicoplanin (VRE Fm Van A), vancomycinresistant Enterococcus faecium sensitive to teicoplanin (VRE Fm Van B),vancomycin resistant Enterococcus faecalis also resistant to teicoplanin(VRE Fs Van A), vancomycin resistant Enterococcus faecalis sensitive toteicoplanin (VRE Fs Van B), enterococcus gallinarium of the Van Agenotype (VRE Gm Van A), enterococcus gallinarium of the Van C-1genotype (VRE Gm Van C-1), enterococcus casseliflavus of the Van C-2genotype (VRE Cs Van C-2), enterococcus flavescens of the Van C-2genotype (VRE Fv Van C-2), and penicillin-sensitive Streptococcuspneumoniae (PSSP) and penicillin-resistant Streptococcus pneumoniae(PSRP). Because of the inability of PSSP and PSRP to grow well inMueller-Hinton broth, MICs with those strains were determined usingeither TSA broth supplemented with defibrinated blood or blood agarplates. Compounds which had significant activity against the strainsmentioned above were then tested for MIC values in a larger panel ofclinical isolates including the species listed above as well asnon-speciated coagulase negative Staphylococcus both sensitive andresistant to methicillin (MS-CNS and MR-CNS). In addition, they weretested for MICs against gram negative organisms, such as Escherichiacoli and Pseudomonas aeruginosa.

2. Determination of Kill Time

Experiments to determine the time required to kill the bacteria wereconducted as described in Lorian, “Antibiotics in Laboratory Medicine”,Fourth Edition, Williams and Wilkins (1991). These experiments wereconducted normally with both staphylococcus and enterococcus strains.

Briefly, several colonies were selected from an agar plate and grown at35° C. under constant agitation until it achieved a turbidity ofapproximately 1.5 and 10⁸ CFU/mL. The sample was then diluted to about6×10⁶ CFU/mL and incubated at 35° C. under constant agitation wascontinued. At various times aliquots were removed and five ten-foldserial dilutions were performed. The pour plate method was used todetermine the number of colony forming units (CFUs).

In general, glycopeptide antibiotics active in the above tests in vitrotests are suitable for use in the pharmaceutical compositions of thisinvention.

B. In Vivo Determination of Antibacterial Activity

1. Acute Tolerability Studies in Mice

In these studies, a pharmaceutical composition of this invention wasadministered either intravenously or subcutaneously and observed for5–15 minutes. If there were no adverse effects, the dose was increasedin a second group of mice. This dose incrementation continued untilmortality occurred, or the dose was maximized. Generally, dosing beganat 20 mg/kg and increased by 20 mg/kg each time until the maximumtolerated dose (MTD) is achieved.

2. Bioavailability Studies in Mice

Mice were administered a compound of this invention either intravenouslyor subcutaneously at a therapeutic dose (in general, approximately 50mg/kg). Groups of animals were placed in metabolic cages so that urineand feces could be collected for analysis. Groups of animals (n=3) weresacrificed at various times (10 min, 1 hour and 4 hours). Blood wascollected by cardiac puncture and the following organs wereharvested—lung, liver, heart, brain, kidney, and spleen. Tissues wereweighed and prepared for HPLC analysis. HPLC analysis on the tissuehomogenates and fluids was used to determine the concentration of thetest compound or IiI present. Metabolic products were also determined atthis juncture.

3. Mouse Septicemia Model

In this model, an appropriately virulent strain of bacteria (mostcommonly S. aureus, or E. Faecalis or E. Faecium) was administered tomice (N=5 to 10 mice per group) intraperitoneally. The bacteria wascombined with hog gastric mucin to enhance virulence. The dose ofbacteria (normally 10⁵–10⁷) was that sufficient to induce mortality inall of the mice over a three day period. One hour after the bacteria wasadministered, a pharmaceutical composition of this invention wasadministered in a single dose either IV or subcutaneously. Each dose wasadministered to groups of 5 to 10 mice, at doses that typically rangedfrom a maximum of about 20 mg/kg to a minimum of less than 1 mg/kg. Apositive control (normally vancomycin with vancomycin sensitive strains)was administered in each experiment. The dose at which approximately 50%of the animals are saved was calculated from the results.

4. Neutropenic Thigh Model

In this model, antibacterial activity of a pharmaceutical composition ofthis invention was evaluated against an appropriately virulent strain ofbacteria (most commonly S. aureus, or E. Faecalis or E. Faecium,sensitive or resistant to vancomycin). Mice were initially renderedneutropenic by administration of cyclophosphamide at 200 mg/kg on day 0and day 2. On day 4 they were infected in the left anterior thigh by anIM injection of a single dose of bacteria. The mice were thenadministered the test compound one hour after the bacteria and atvarious later times (normally 1, 2.5, 4 and 24 hours) the mice weresacrificed (3 per time point) and the thigh excised, homogenized and thenumber of CFUs (colony forming units) were determined by plating. Bloodwas also plated to determine the CFUs in the blood.

5. Pharmacokinetic Studies

The rate at which a compound of this invention is removed from the bloodcan be determined in either rats or mice. In rats, the test animals werecannulated in the jugular vein. The test compound was administered viatail vein injection, and at various time points (normally 5, 15, 30,60minutes and 2,4,6 and 24 hours) blood was withdrawn from the cannula. Inmice, the test compound was also administered via tail vein injection,and at various time points. Blood was normally obtained by cardiacpuncture. The concentration of the remaining test compound wasdetermined by HPLC.

In general, the pharmaceutical compositions of this invention wereactive in the above test in vivo and demonstrated a broad spectrum ofactivity.

Example 5 Determination of Tissue Accumulation

A. Tissue Distribution Using Radiolabeled Compound

This procedure is used to examine the tissue distribution, excretion andmetabolism of a radiolabeled test compound in both male and female ratsfollowing intravenous infusion at 10 mg/kg. Male and femaleSprague-Dawley rats (n=2 per sex per compound) are dosed with ³H-labeledtest compound at 10 (400 μCi/kg) and 12.5 mg/kg (100 μCi/kg),respectively, via intravenous infusion (˜2 min). The test compound isformulated in 5% hydroxypropyl-β-cyclodextrin as 2.5 mg/mL solution.Urine and feces are cage collected over 24 hours period. At 24 hoursafter dosing, animals are sacrificed and tissues are removed. Serum,urine and tissues are analyzed for total radioactivity by oxidationfollowed by liquid scintillation counting. Urine and selected tissuessamples are extracted and analyzed by reverse phase HPLC withradioactive flow detector for the presence of potential metabolites.

B. Tissue Accumulation Following Single Dose

This procedure is used to evaluate tissue distribution of a testcompound in rats following single dose administration by infusion. MaleSprague-Dawley rats (n=3 per dose groups) are dosed with 50 mg/kg of atest compound. Two formulations are used: 30% PEG 400 and 10%sulfobutylether-β-cyclodextrin. Urine samples are cage collected over 24hours. Blood samples are collected for serum chemistry and concentrationdetermination. Liver and kidneys are removed for histology evaluation.One kidney and part of the liver are homogenized for concentrationanalysis using reverse phase HPLC with UV detection. Drug concentrationsin urine and serum samples are determined by LC-MS analysis.

C. Tissue Distribution Following Multiple Doses

This procedure is used too evaluate the potential tissue accumulation ofa test compound in rats following multiple dose administration byintravenous infusion. Male and female Sprague-Dawley rats (n=4 per sexper dose group) are dosed with a test compound at 12.5, 25 and 50 mg/kgper day for seven days. Animals are sacrificed at day 1 (n=3 per sex perdose group) following the last dose administered. One animal per sex perdose group is retained as recovery animal and sacrificed at day 7following the last dose administered. The test compound is formulated in5% hydroxypropyl-β-cyclodextrin or 1% sucrose/4.5% dextrose. Urinesamples are cage collected at days 1 and 7 post-dose. Blood samples arecollected for serum chemistry and concentration determination. Liver andkidneys are removed for histology evaluation. One kidney and part of theliver are homogenized for concentration analysis using reverse phaseHPLC with UV detection. Drug concentrations in urine and serum samplesare determined by LC-MS analysis.

Nephrotoxicity, histamine release and vascular irritation can bemeasured using procedures well-known to those skilled in the art.

Example 6 Determination of Tissue Accumulation and Nephrotoxicity

Female Sprague-Dawley rates (three per group) were dosed with Compound A(50 mg/kg) formulated in either hydroxypropyl-β-cyclodextrin or 5%dextrose/water or vehicle. The indicated doses were administered in avolume of 10 mL/kg via a 2-minute intravenous infusion. At 24 hours, theanimals were sacrificed and serum and tissue samples collected. Theresults are shown in Tables 1 and 2.

TABLE 1 Tissue Distribution and Urinary Recovery for Compound A inVarious Formulations Following Intravenous Infusion to Female Rats at adose of 50 mg/kg. (Values are Mean (SD)) Serum % Recovered CompoundConc. (as unchanged parent) (Formulation) (μg/mL) Urine Liver KidneyCompound A 0.86 (0.19) 90.91 (8.39)  1.90 (0.32)  0.62 (0.12) (25% CD)Compound A 1.66 (0.33) 40.51  4.89 (0.81)  2.08 (0.43) (5% CD) (18.57)Compound A 17.1 (12.1) 17.45 (6.92)  8.47 (0.46)  5.68 (2.49) (1% CD)Compound A 59.8 (27.1) 12.61 (4.60) 14.19 (3.41) 17.82 (4.94) (D5W) CD =hydroxypropyl-β-cyclodextrin D5W = aqueous 5% dextrose solution

As shown in Table 1, urinary recovery of Compound A was significantlyhigher in formulations contain a cyclodextrin; and liver and kidneyaccumulation were significantly lower in such formulations.

TABLE 2 Effects of Compound A Formulation on Serum Renal ChemistryCompound Formulation BUN (mg/dL) Creatinine (mg/dL) Vehicle 25% (w/v) CD14 ± 1 0.26 ± 0.06 Compound A 25% (w/v) CD 13 ± 2 0.26 ± 0.02 Vehicle 5% (w/v) CD 10 ± 1 0.21 ± 0.01 Compound A  5% (w/v) CD 18 ± 5 0.31 ±0.07 Vehicle  1% (w/v) CD 13 ± 2 0.24 ± 0.01 Compound A  1% (w/v) CD 26± 5 0.34 ± 0.08 Vehicle D5W 12 ± 2 0.28 ± 0.02 Compound A D5W 67 ± 20.72 ± 0.08 CD = hydroxypropyl-β-cyclodextrin D5W = aqueous 5% dextrosesolution

The results in Table 2 show that the formulations containingcyclodextrin had significantly less nephrotoxicity compared toformulations without cyclodextrin.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto. Additionally, all publications, patents, andpatent documents cited hereinabove are incorporated by reference hereinin full, as though individually incorporated by reference.

1. A method for reducing nephrotoxicity produced by a lipidatedglycopeptide antibiotic in a mammal, the method comprising administeringa therapeutically effective amount of the lipidated glycopeptideantibiotic or a pharmaceutically acceptable salt thereof to the mammalin a pharmaceutical composition comprising a cyclodextrin, wherein thelipidated glycopeptide antibiotic produces nephrotoxicity in the mammalin the absence of the cyclodextrin.
 2. The method of claim 1, whereinthe pharmaceutical composition further comprises water.
 3. A method forreducing nephrotoxicity produced by a lipidated glycopeptide antibioticin a mammal, the method comprising administering a therapeuticallyeffective amount of the lipidated glycopeptide antibiotic or apharmaceutically acceptable salt thereof to the mammal in apharmaceutical composition comprising: (a) an aqueous cyclodextrincarrier, wherein the lipidated glycopeptide antibiotic producesnephrotoxicity in the mammal in the absence of the cyclodextrin.
 4. Amethod for reducing nephrotoxicity produced by a lipidated glycopeptideantibiotic in a mammal, the method comprising administering atherapeutically effective amount of the lipidated glycopeptideantibiotic or a pharmaceutically acceptable salt thereof to the mammalin a pharmaceutical composition comprising: (a) 1 to 40 weight percentof a cyclodextrin; and 60 to 99 weight percent of water, based on 100weight percent of the composition, wherein the lipidated glycopeptideantibiotic produces nephrotoxicity in the mammal in the absence of thecyclodextrin.
 5. The method of claim 4, wherein the cyclodextrincomprises about 5 to 35 weight percent of the composition.
 6. The methodof claim 4, wherein the cyclodextrin comprises about 10 to 30 weightpercent of the composition.
 7. The method of any one of claims 1 to 6,wherein the cyclodextrin is hydroxypropyl-β-cyclodextrin.
 8. The methodof any one of claims 1 to 6, wherein the cyclodextrin is sulfobutylether β-cyclodextrin.